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CRISPR/Cas9
Published in Sylvia Uzochukwu, Nwadiuto (Diuto) Esiobu, Arinze Stanley Okoli, Emeka Godfrey Nwoba, Christpeace Nwagbo Ezebuiro, Charles Oluwaseun Adetunji, Abdulrazak B. Ibrahim, Benjamin Ewa Ubi, Biosafety and Bioethics in Biotechnology, 2022
Sylvia Uzochukwu, Arinze S. Okoli
Gene editing involves deletion, insertion or modification of specific DNA sequences in a genome by modified enzymes with great precision. Although CRISPR/Cas9 has become synonymous with gene editing, restriction enzymes have initially been applied to edit DNA. However, restriction enzyme-mediated gene editing was neither precise nor targeted. The possibility of targeted gene editing came about with the discovery of zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Both ZFN and TALENs involve engineering an enzyme with two domains: DNA-binding domain and a restriction endonuclease domain. The DNA-binding domain targets and binds to specific DNA sequence on the genome, and the endonuclease domain cleaves the DNA at the desired site. Although ZFNs and TALENs can make targeted changes to the genome, they are both inefficient, restricted to certain cell types, very expensive and cumbersome to produce and apply. The CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/associated protein 9), which can be applied in all cell types, is based on interactions between DNA, RNA and protein. The system offers a far simpler, cheaper, efficient and easier way of editing genes and has become the popular and preferred way of editing the genome of an organism (Okoli et al. 2018).
The genetically modified food credibility gap
Published in Charlotte Fabiansson, Stefan Fabiansson, Food and the Risk Society, 2016
Charlotte Fabiansson, Stefan Fabiansson
Genetically engineered plants are generated in a laboratory by altering their genetic makeup and are initially tested in the laboratory for desired qualities before undergoing field trials and possible commercialisation. Genetic modification can involve either the insertion or deletion of genes to create a genetically modified organism. When genes are inserted, they usually come from a different species, which is a form of horizontal gene transfer. The techniques used might involve attaching the genes to a virus that penetrate the recipient cell. Additional methods include the physical insertion of extra DNA into the nucleus of the intended host with a very small syringe, using electroporation by introducing DNA from one organism into the cell of another by use of an electric pulse, or with very small particles fired from a particle gun; the biolistic method. Some methods exploit natural forms of gene transfer, such as the ability of Agrobacterium tumefaciens to transfer genetic material to plants, or the ability of lentiviruses to transfer genes to animal cells. Most GM plants have been generated by the biolistic method or by Agrobacterium tumefaciens mediated transformation (Khan and Liu 2009).
Applications of Antiviral Nanoparticles in Cancer Therapy
Published in Devarajan Thangadurai, Saher Islam, Charles Oluwaseun Adetunji, Viral and Antiviral Nanomaterials, 2022
Anusha Konatala, Sai Brahma Penugonda, Fain Parackel, Sudhakar Pola
The replacement or modification of the defective genes with the healthy version of the gene is called gene therapy. Cancer as a disease also has a genetic aspect to it. It is driven by the overexpression or inhibition of certain genes (for example, the inhibition of tumour-suppressing genes). Earlier, there were concerns about exploring gene therapy for cancer treatment among the scientific community. In recent years, gene therapy has been among the most promising treatments for challenging diseases globally. A threat in traditional cancer treatment is multi-drug resistance (MDR). This resistance is caused by the repeated influx of proteins on the cell membrane, and, as a consequence, the therapeutic efficacy of the drug is depleted (Quader and Kataoka 2017). Most commonly in patients with MDR, unexpected results due to traditional therapies are reported. These hurdles can be mitigated with the use of gene therapy in combination with the drugs. Li et al. studied cancer treatment for non-small-cell lung cancer (NSCLC), caused mostly due to a mutated P53 gene by the advantage of the combination therapy of p53 gene and bortezomib (BTZ). This combined treatment was masterfully used with mesoporous hollow silica nanospheres (Quader and Kataoka 2017). The reports explained the refurbishment and revitalisation of the p53 signalling pathway. Xu et al. studied the pigment epithelium derived factor (PEDF) in combination with paclitaxel (PTX) to inhibit cancer development in a colon cancer C26 subcutaneous model using PEG-Poly (lactic-co-glycolic acid) (PLGA) NP-based co-delivery system. The reports identified aggregate PEDF expression, high apoptosis, depleted angiogenesis, and, importantly, G2/M cell-cycle phase breakdown (Xu et al. 2016).
Are attitudes toward labeling nano products linked to attitudes toward GMO? Exploring a potential ‘spillover’ effect for attitudes toward controversial technologies
Published in Journal of Responsible Innovation, 2019
Heather Akin, Sara K. Yeo, Christopher D. Wirz, Dietram A. Scheufele, Dominique Brossard, Michael A. Xenos, Elizabeth A. Corley
Genetic modification involves humans intentionally changing an organism’s genetic material to add or alter its characteristics (National Academies of Sciences 2016). Biologists began using genetic modification in the 1980s and as a few select and widely grown crops were commercialized, it became a topic of public interest. In the U.S., the debate over labeling genetically modified organisms rose to prominence in the media in the late 1990s (Shanahan, Scheufele, and Lee 2001). Initially, opinion polls indicated the majority of publics supported labeling GMOs, although attitudes varied substantially by country (Shanahan, Scheufele, and Lee 2001). This yielded different regulatory and labeling policies across the world, reflecting the level of public support or concern for these technologies within each nation or region (Caswell 2000). For instance, aversion to GMO products was relatively high throughout Europe, which incited mandatory labeling, while North American consumers were less concerned and so agencies did not initially require labeling (Fulton and Giannakas 2004). In the U.S., the debate became a national issue again in 2016 when Congress passed a bill requiring labeling of foods using genetically-engineered ingredients (Strom 2016). Despite these differential public responses to GM worldwide, the scientific consensus suggests that GM foods are no more risky to consume than conventional foods (National Academies of Sciences 2016).
Development of capability for genome-scale CRISPR-Cas9 knockout screens in New Zealand
Published in Journal of the Royal Society of New Zealand, 2018
Francis W. Hunter, Peter Tsai, Purvi M. Kakadia, Stefan K. Bohlander, Cristin G. Print, William R. Wilson
Few New Zealand scientists will be unaware of CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9) as a remarkable new technology that is transforming many aspects of biological research. This ancient apparatus of adaptive immunity, widely distributed in archaea and bacteria (Bhaya et al. 2011), is based on the recognition of foreign nucleic acids rather than peptides, facilitating its adaptation as a molecular biology tool. CRISPR-Cas systems have been reengineered to seek out and manipulate genes in living cells (including higher eukaryotes) providing techniques for precise re-writing of DNA sequences (gene editing) (Komor et al. 2016; Paquet et al. 2016), inactivating genes (gene knockout) (Kleinstiver et al. 2016), transcriptional or epigenetic modification of levels of gene expression (Larson et al. 2013; Perez-Pinera et al. 2013), dynamic imaging of genomic loci and RNA movement in cells (Chen et al. 2013) and as a diagnostic for facile and extremely sensitive detection of specific nucleic acids (Gootenberg et al. 2017). These tools are already finding widespread and diverse application in the biological and biomedical sciences in New Zealand and may yet play an important role in our biosecurity through, for example, CRISPR-mediated gene drive technology (Hammond et al. 2015).
Gene Editing: A View Through the Prism of Inherited Metabolic Disorders
Published in The New Bioethics, 2018
The ‘technology of purposeful DNA modification’ (Rehmann-Sutter 2018), that is the ability deliberately to make targeted alterations in specific genes within an organism, has been developed to such a sophisticated level that gene editing is now a clinical therapeutic reality. Stem cells extracted from patients affected with severe paediatric neurodegenerative disorders such as metachromatic leukodystrophy have been edited ex vivo using lentiviral systems to insert a correctly functioning copy of the defective gene, and the cells returned to the patients as an autologous haematopoietic stem cell transplant, thereby preventing the progression of the disease (Biffi et al. 2013). More recently in vivo gene editing has been accomplished using zinc finger nuclease technologies (Sharma et al. 2015) to insert a functioning copy of the gene encoding specific lysosomal enzymes directly in to liver cells of patients with lysosomal storage disorders in an attempt to cure their disease, with the first patients treated widely reported in the press.1,2 The development of RNA-guided gene-editing technologies such as CRISPR/Cas93 or zinc finger nucleases means that it is now possible to edit single genes within an organism, including an embryo.